Perchlorate (${ClO_4}^-$) is an emerging contaminant found in surface water and soil/groundwater. Microbial removal of perchlorate is the method of choice since perchlorate-reducing bacteria (PRB) can reduce perchlorate to harmless end-products. A previous study [3] showed experimental evidence of autotrophic perchlorate removal using elemental sulfur granules and activated sludge. The granular sulfur is a relatively inexpensive electron donor, and activated sludge is easily available from a wastewater treatment plant. A batch test was performed in this study to further investigate the effect of various environmental parameters on the perchlorate degradation by sludge microorganisms when elemental sulfur was used as electron donor. Results of the batch test suggest optimum conditions for autotrophic perchlorate degradation by sludge microorganisms. The results also show that sulfur-oxidizing PRB enriched from activated sludge removed perchlorate better than activated sludge. Taken together, this study suggests that autotrophic perchlorate removal using elemental sulfur and activated sludge can be improved by employing optimized environmental conditions and enrichment culture.